Journal: Journal of Endocrinology

Article Title: Local adiponectin treatment reduces atherosclerotic plaque size in rabbits

doi: 10.1677/joe-06-0173

Figure Lengend Snippet: Figure 1 Serum adiponectin concentrations in rabbits. 1, Aorta non-injured rabbits on normal cholesterol diet; 2, aorta injured rabbits on high-cholesterol diet before adenovirus transfer; 3, after transfer Ad-bgal through intima; 4, after transfer Ad-APN through intima; 5, after transfer Ad-bgal through adventitia; 6, After transfer Ad-APN through adventitia. The symbol * indicates significant difference (P!0.01) vs serum adiponectin concentrations in aorta non-injured rabbits on normal cholesterol diet; † compared with rabbits before adenovirus transfer; and ‡ compared with rabbits after Ad-bgal transfer. The values are meansGS.E.M.

Article Snippet: Following blocking with 5% non-fat milk, the blots were incubated with anti-His tag mouse monoclonal antibody (1:1000 dilution, Applygen Technologies Inc., Beijing, China) and rabbit adiponectin polyclonal antibody (1:100 dilution, Boster, Wuhan, Hubei, China) at 4 8C overnight.

Techniques:

Journal: Journal of Endocrinology

Article Title: Local adiponectin treatment reduces atherosclerotic plaque size in rabbits

doi: 10.1677/joe-06-0173

Figure Lengend Snippet: Figure 2 SDS-PAGE (A) and western blot analysis of the supernatant of cultured Pichia pastoris (B, using anti-His tag monoclonal antibody; C, using adiponectin polyclonal antibody). Line 1 is a blank pGAPZa Pichia vector (not containing adiponectin gene) sample control and lines 2–4 are samples from the supernatant of cultured Pichia pastoris containing the recombinant rabbit adiponectin gene. Line 5 contained the molecular weight markers (not visible in image of blot). Both anti-His antibody and adiponectin polyclonal antibody detected a single predominant protein at z42 kDa (containing 9.3 kDa a-factor signal peptide and 2.5 kDa C-terminal His-tag).

Article Snippet: Following blocking with 5% non-fat milk, the blots were incubated with anti-His tag mouse monoclonal antibody (1:1000 dilution, Applygen Technologies Inc., Beijing, China) and rabbit adiponectin polyclonal antibody (1:100 dilution, Boster, Wuhan, Hubei, China) at 4 8C overnight.

Techniques: SDS Page, Western Blot, Cell Culture, Plasmid Preparation, Control, Recombinant, Molecular Weight

Journal: Journal of Endocrinology

Article Title: Local adiponectin treatment reduces atherosclerotic plaque size in rabbits

doi: 10.1677/joe-06-0173

Figure Lengend Snippet: Figure 5 Effect of Ad-APN treatment on adhesion molecules in atherosclerotic rabbits 14 days after recombinant adenovirus transfer. A, Immunohistochemical detection of adiponectin, VCAM-1 and ICAM-1 in atherosclerotic lesions. Representative images are shown. 1, Ad-bgal transfer through intima. 2, Ad-APN transfer through intima. 3, Ad-bgal transfer through adventitia. 4, Ad-APN transfer through adventitia. The magnification is 400. B, mRNA expression of VCAM-1 and ICAM-1 in abdominal aortic tissue in Ad-bgal or Ad-APN-treated rabbits. The values are meansGS.E.M.

Article Snippet: Following blocking with 5% non-fat milk, the blots were incubated with anti-His tag mouse monoclonal antibody (1:1000 dilution, Applygen Technologies Inc., Beijing, China) and rabbit adiponectin polyclonal antibody (1:100 dilution, Boster, Wuhan, Hubei, China) at 4 8C overnight.

Techniques: Recombinant, Immunohistochemical staining, Expressing

Journal: The Journal of Clinical Investigation

Article Title: MicroRNA-132 enhances transition from inflammation to proliferation during wound healing

doi: 10.1172/JCI79052

Figure Lengend Snippet: (A) Representative images of wounds on the backs of miR-132–KO mice (n = 9) and their WT littermate controls (n = 6) on days 0, 2, 4, 6, and 8 after wounding. Wound closures were quantified and are presented as the percentage of the initial wound area size. (B) qRT-PCR was performed to detect Ccl20, Cxcl5, Cxcl1, Tnf, Il1a, and Il1b expression levels in the intact and wound-edge skin of both WT (n = 5) and KO (n = 9) mice 2 days after injury. *P < 0.05 and **P < 0.01 by Student’s t test. (C) H&E staining (left panels) and immunostaining of Gr-1 (middle and right panels) in the wound area in WT and KO mice 2 days after injury. White arrows demarcate wound edges. Scale bars: 50 μm. (D) Chemotaxis of neutrophils isolated from the peripheral blood of WT (n = 5) and KO mice (n = 7) toward the medium containing 8 μM N-formylmethionyl-leucyl-phenylalanine (fMLP). Migrating cells were quantified by flow cytometry. Plots show the FSC/SSC of the migrated cells. (E) Immunostaining of Ki-67 and HB-EGF in wound-edge skin from WT (n = 9) and KO mice (n = 5) 2 days after injury. Sections were counterstained using DAPI (blue, nucleus). The number of Ki-67–positive cells was counted. (F) Immunostaining of p-STAT3 and p-ERK in wound-edge skin from WT (n = 9) and KO mice (n = 5) 2 days after injury. Scale bars: 50 μm (E and F). *P < 0.05, **P < 0.01, and ***P < 0.001 by Student’s t test.

Article Snippet: Neutrophils, macrophages, and T cells were detected using rabbit anti-mouse Gr-1 (1:100; catalog 550291; BD Pharmingen); rabbit anti-mouse CD68 (1:10,000; catalog MCA1957; AbD Serotec); and rabbit anti-mouse CD4 (1:100; catalog 553647; BD Pharmingen).

Techniques: Quantitative RT-PCR, Expressing, Staining, Immunostaining, Chemotaxis Assay, Isolation, Flow Cytometry